The Corynebacterium pyogenes infection in cattle. 1. Incidence of the pathogen]

Reported are the incidence of Corynebacterium pyogenes together with different pathological changes as well as the existence of latent Corynebacterium pyogenes infections and their widespread occurrence. Corynebacterium pyogenes was established from 609 in 2,130 samples of pathological processes, accounting for 28.6%. The pathogen was cultivated from various processes, including enlarged tail lymph nodes (61.1%), tail phlegmons (56.3%), abscesses (49.1%), epiphysiolyses (45.2%), liver abscesses (31.8%), panaritia at beginning of fattening (20.5%), aborted foetuses (14.9%), foetal membranes in cases of incarcerated placenta (12.0%), and panaritia on end of fattening (3.4%). The same pathogenic microorganism was recorded from nine per cent of apparently intact heifer udders, before pasturing. Corynebacterium pyogenes was cultivated also from nasal mucous membrane (8.4%) and retropharyngeal lymph nodes (37.2%). The highest detection rate was 71.6%, obtained from the tonsils.     

Effect of damage to the teat end on the experimental induction of mastitis in dry cows with Corynebacterium pyogenes

The teat ends of 12 dry cows were contaminated with Corynebacterium pyogenes. To determine whether a pre-existing (an)aerobic bacterial infection of the udder was a predisposing factor for a C pyogenes mastitis they included infected and uninfected quarters. Anaerobic bacteria could not be found and mastitis was not induced. When the teats were contaminated with C pyogenes after the teat ends had been injured 30 of the quarters became infected, and anaerobic bacteria were demonstrated in many quarters.     

Corynebacterium pyogenes mastitis among heifer calves.

In August 1975 an outbreak of C pyogenes mastitis occurred involving 18 heifer calves at the National Institute for Research in Dairying. This paper describes the outbreak which affected animals varying in age from five to 22 months. Fourteen of the cases (77%) were among calves less than 10 months old. The advisability of using fly control measures among young stock in the summer months is discussed.     

Carriage of Corynebacterium pyogenes by the cattle nuisance flies Hydrotaea irritans (Fallén) and Musca autumnalis (De Geer)

Two species of cattle-visiting Muscidae were experimentally contaminated with C. pyogenes, a pathogen involved in the aetiology of summer mastitis. Surface contamination persisted for at least 4 days. Since M. autumnalis would not feed on media containing C. pyogenes the bacterium did not persist internally. All C. pyogenes were eliminated from the gut of H. irritans in 4 days. H. irritans is thus more likely to transmit C. pyogenes than is M. autumnalis but only by mechanical transfer, and is not a true vector.     

Infection following challenge of the lactating and dry udder of dairy cows with Actinomyces pyogenes and Peptostreptococcus indolicus

Challenge of 12 mammary glands of cows in mid-lactation with 10(7) colony forming units (cfu) of Peptostreptococcus indolicus on two occasions led to clinical mastitis in only four quarters. The bacteria were rarely recovered and disappeared from the secretion within 14 days. In challenges 7 days prior to drying off eight of 12 quarters became infected and at drying off all quarters challenged became infected. The infections established at drying off persisted well into the dry period. P. indolicus infection was also established in all of 12 dry glands challenged, but usually eliminated at calving or early in the next lactation. Isolation of P. indolicus was accompanied in about one-third of cases by changes in the appearance of the secretion. Intramammary challenge with Actinomyces (formally Corynebacterium) pyogenes led to clinical and subclinical infections in nine of 12 lactating glands and in all of six dry glands. Dry period infections with A. pyogenes were more severe and rarely eliminated even by antibiotic therapy. Infections during lactation were often eliminated either naturally or by antibiotic therapy. Intermittent recovery of A. pyogenes from the lactating mammary gland, without clinical signs of infection, was possible for up to 90 days after challenge. Combined infections with A. pyogenes and P. indolicus were clinically more severe with a higher frequency of systemic involvement. It was shown that in the non-lactating gland an acute mastitis, similar to 'summer mastitis' could be established either by simultaneous inoculation with A. pyogenes and P. indolicus or by subsequent inoculation of quarters excreting P. indolicus with A. pyogenes.     

Feline pyothorax caused by a Borrelia-like organism and Corynebacterium pyogenes.

A 2-year-old, neutered, male cat was euthanatized because of difficult repiration and inability to rise. Necropsy revealed pyothorax. Corynebacterium pyogenes was grown from the purulent thoracic exudate. A Borrelia-like microorganism was demonstrated in the exudate, using phase-contrast microscopy.     

Cause, occurrence, and clinical signs of mastitis and anorexia in cows in a Wisconsin study

Microbiologic culture revealed the following cause of mastitis and anorexia in 145 cows in Wisconsin to be Escherichia coli, 66 cows; Klebsiella spp, 3; Corynebacterium pyogenes, 27; streptococci, 21; staphylococci, 20; yeasts, 1; and no bacterial growth, 7. Mastitis was detected with approximately equal frequency throughout the year. Escherichia coli was isolated throughout the year, but was more common and was the predominant organism during the summer. Corynebacterium pyogenes was isolated most often in winter and spring; streptococci in fall, winter, and spring; and staphylococci throughout the year. Corynebacterium pyogenes caused most of the mastitis in nonlactating cows. Escherichia coli, C pyogenes, streptococci, and staphylococci were isolated with about equal frequency at parturition, whereas E coli was the predominant cause of mastitis in early and late lactation. Of cases of mastitis, 27% were seen 10 days before and after parturition. Local and systemic clinical signs of infection were similar for all causes, except that C pyogenes caused more (P less than 0.01) malodorous and purulent milk than did other organisms and was isolated more commonly from quarters with injured teats. Recovery was significantly (P less than 0.01) higher in cows with E coli infections, compared with recovery in cows with gram-positive organism infections. Cows with C pyogenes infections frequently had quarters that ultimately ceased lactation. A few cows were recumbent at initiation of antimicrobial therapy and a few were not eating 24 hours later; however, 50% of these cows recovered. Criteria such as season of year, stage of lactation, appearance of milk and udder, and appetite permitted the cause (gram-negative or gram-positive organisms) of the mastitis to be predicted with 77% accuracy.     

Failure to induce in rabbits effective immunity to a mixed infection of Fusobacterium necrophorum and Corynebacterium pyogenes with a combined bacterin.

Failure to induce in rabbits effective immunity to a mixed infection of Fusobacterium necrophorum and Corynebacterium pyogenes with a combined bacterin. Onderstepoort Journal of Veterinary Research, 44 (4), 253--2;6 (1977). Rabbits were immunized with alum-precipitated, oil adjuvant and an untreated bacterin composed of F. necrophorum and C. pyogenes. Immunized rabbits were challenged intradermally with a mixture of F. necrophorum and C. pyrogenes. Immunized rabbits were challenged intradermally with a mixture of F. necorphorum and C. pyrogenes. Initially a low level of initial transient resistance could be demonstrated but a solid immunity could not be established.     

Immunofluorescence tests for the detection of antibodies against Corynebacterium pyogenes and streptococci in blood serum and vaginal mucus of cattle]

Bacteriological tests were applied to cattle with endometritis and vaginitis. Included were cervical mucus samples and immunofluorescence tests to detect in that mucus as well as in blood serum antibody to Corynebacterium pyogenes and Streptococcus haemolyticus. The results pointed at intensive contact of the animals with the above pathogens and to their frequent occurrence in cervical mucus of cattle afflicted with endometritis and vaginitis. They also supported the assumption of localised antibody formation in the sexual organs or female cattle.     

Comparative bacteriological studies on summer mastitis in grazing cattle and pyogenes mastitis in stabled cattle in Denmark.

A total of 143 secretions from clinical cases of summer mastitis (SM) in grazing cattle and from 89 cases of pyogenes mastitis (PM) in stabled cattle were examined bacteriologically. The typical bacteriological finding was a mixed flora in which the predominant organisms were Actinomyces pyogenes (SM-70%, PM-85%), Peptostreptococcus indolicus (54%, 54%), a microaerophilic coccus (Stuart-Schwan coccus) (26%, 25%), Fusobacterium necrophorum biovar B (22%, 12%), Bacteroides melaninogenicus (20%, 9%) and Streptococcus dysgalactiae (21%, 5%). All except six cases occurred in non-lactating animals or within three weeks after parturition. The majority of animals (about 90%) had only one quarter affected and no differences in quarter distribution were observed between the two groups.     

Isolation of Corynebacterium pilosum and Actinomyces pyogenes from cystitis and vulvovaginitis infection in a 2-month-old female calf

Cystitis and vulvovaginitits, due to Corynebacterium pilosum and Actinomyces pyogenes infection in a 2-month-old female calf, is described. The prominent clinical signs were urinary incontinence, adherence of triple phosphate crystals to the vulvar hair and ulceration on the vulva, the ventral side of the tail skin and the perineum. Only a mild inflammation of the bladder mucosa and submucosa was seen on histological examination.     

Rapid identification of bovine mastitis pathogens by high-resolution melt analysis of 16S rDNA sequences

Accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. Here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (HRMA) of 16S rDNA sequences. Real-time PCR amplification of 16S rRNA gene fragment, spanning the variable region V5 and V6 was performed with a resulting amplicon of 290bp. First, a library was generated of melt curves of 9 common pathogens that are implicated in bovine mastitis. Six of the isolates, Escherichia coli, Streptococcus agalactiae, Klebsiella pneumoniae, Streptococcus uberis, Staphylococcus aureus and Mycoplasma bovis, were type strains while the other 3, Arcanobacterium pyogenes, Corynebacterium bovis and Streptococcus dysgalactiae, were bovine mastitis field isolates. Four of the type strains, E. coli, S. agalactiae, K. pneumoniae and S. aureus, were found to be of human origin, while the other 3 type strains were isolated from bovine infections. Secondly, the melt curves and corresponding amplicon sequences of A. pyogenes, E. coli, S. agalactiae, S. dysgalactiae, K. pneumoniae, S. uberis and S. aureus were compared with 10 bovine mastitis field isolates of each pathogen. Based on the distinct differences in melt curves and sequences between human and bovine isolates of E. coli and K. pneumoniae, it was deemed necessary to select a set of bovine strains for these pathogens to be used as reference strains in the HRMA. Next, the HRMA was validated by three interpreters analyzing the differential clustering pattern of melt curves of 60 bacterial cultures obtained from mastitis milk samples. The three test interpreters were blinded to the culture and sequencing results of the isolates. Overall accuracy of the validation assay was 95% as there was difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of S. uberis. The present study revealed that broad-range real-time PCR with HRMA can be used as a powerful, fast and low-cost tool for the differentiation of clinically important bacterial mastitis pathogens.     

Corynebacterium pyrogenes; a biochemical and serological study

The study comprises 136 strains of Corynebacterium pyogenes originating from cattle (105), swine (20), sheep (1), and insects (10). For comparison 2 strains of human origin and 1 strain of Gorynebacterium hemolyticum were examined.One of the bovine strains was atypical, being gelatinase-negative, otherwise the strains of Cb. pyogenes were found to be biochemically identical apart from minor deviations in fermentation patterns (Table 1). Neither were antigenic differences demonstrated (gel diffusion analyses, Figs. 1 and 2).Both of the human strains agreed biochemically with Cb. pyogenes (Table 1). By gel diffusion cross analyses one of them was found to be identical with Cb. pyogenes, the other not, though anti-genically related to it (Fig. 2).Gb. hemolyticum deviated biochemically as well as serologically from Gb. pyogenes, but the 2 organisms shared antigenic determinants (Fig. 3).     

Experimental Corynebacterium pseudotuberculosis mastitis in dairy cows

The inoculation of 2000 colony-forming units of Corynebacterium pseudotuberculosis into one teat canal of each of three cows resulted in severe, chronic, pyogranulomatous mastitis. Within three days the cows had a reduced haematocrit, haemoglobin concentration and red cell count. The anaemia was initially normocytic, normochromic and non-regenerative, and was associated with a brief peak of neutrophilia; a regenerative response became evident two to three weeks later. Clinical signs of mastitis appeared seven to 14 days after the inoculation, with a peak of high fever, more severe anaemia, a second peak of neutrophilia and the complete cessation of milk production from all quarters; extensive and severe pyogranulomatous mastitis developed in the inoculated quarters. No other lesions were detected postmortem, and C pseudotuberculosis was cultured from the affected quarters but not from the supramammary lymph nodes and viscera.     

利血平对化脓隐秘杆菌大环内酯类外排基因mefA表达的影响

化脓隐秘杆菌(Trueperella pyogenes,T.pyogenes)是一种能够引起动物和人化脓性感染的重要病原菌.随着抗生素的广泛使用,该菌已对临床常用的大环内酯类、四环素类等药物产生不同程度的耐药性.本试验拟探讨外排泵抑制剂利血平对T.pyogenes大环内酯类外排基因mefA mRNA及蛋白表达的影响,将...     

Summer mastitis in heifers: studies on the seasonal occurrence of Actinomyces pyogenes, Peptostreptococcus indolicus and Bacteroidaceae in clinically healthy cattle in Denmark.

With the aim of investigating the seasonal occurrence of Actinomyces pyogenes, Peptostreptococcus indolicus, Bacteroides melaninogenicus ss. levii and Fusobacterium necrophorum, and thus the potential for development of summer mastitis, clinically healthy Danish Holstein-Friesian heifers due to calve in the autumn were sampled from the teat tip, the conjunctiva and the oral cavity at 2-6 week intervals from 1979 to 1981. The overall isolation rates of F. necrophorum, P. indolicus and B. melaninogenicus ss. levii, in order of significance, were significantly higher during the pasture period whereas no differences in isolation rates of A. pyogenes between housed and pastured animals were detected. F. necrophorum was recovered almost exclusively from the oral cavity, P. indolicus and A. pyogenes occurred most frequently in samples from the teat skin, whereas isolates of B. melaninogenicus ss. levii were evenly distributed between conjunctiva and teat tip samples. A distinct seasonal pattern of the isolation rates of summer mastitis pathogens was recorded, which corresponded closely to the seasonal activity of symbovine insects, in particular the headfly Hydrotaea irritans (Fallén). However, the high proportion of clinically healthy bacterial carriers as compared with the incidence of clinical disease strongly suggests that as yet unknown contributing or triggering factors, apart from the mere presence of the relevant bacterial species, are required for the establishment and development of clinical summer mastitis.     

Isolation of Arcanobacterium (Actinomyces) pyogenes from cases of feline otitis externa and canine cystitis.

Arcanobacterium pyogenes is a normal inhabitant of the mucous membranes of domestic animals, such as cattle, sheep, swine, and goats. It is also an opportunistic pathogen in these animals, where it causes a variety of purulent infections involving the skin, joints, and visceral organs. Two recent cases of isolation of A. pyogenes from companion animals are reported. In the first case, a cat presented with a chronic otitis externa, from which A. pyogenes was isolated in pure culture. The second case involved a dog with a urinary tract infection, where A. pyogenes was isolated from urine as the predominant bacterial species. In both cases, the A. pyogenes isolates were presumptively identified by macrobiochemical tests, and then their identities were confirmed by polymerase chain reaction analysis and 16S rRNA gene sequencing.     

Experimental infection of lactating bovine mammary glands with Streptococcus uberis in quarters colonized by Corynebacterium bovis

Twenty-seven quarters of 18 lactating dairy cows were inoculated intramammarily with 3.6 X 10(4) colony-forming units (CFU) of a strain of Streptococcus uberis isolated from a cow with clinical mastitis. Before quarters were inoculated, 22 were considered as naturally colonized with Corynebacterium bovis, and 5 were considered bacteriologically negative. Streptococcus uberis was isolated from all quarters within 2 days after inoculation, and all quarters developed clinical mastitis by 3 days after inoculation. Mastitis was acute, and most cows had increased rectal temperatures. The number of somatic cells increased significantly (P less than 0.05), and milk production decreased significantly. In many cows, rectal temperatures remained increased, and Str uberis was isolated from infected glands after intramammary and systemic antimicrobial treatments were given. A decreased number (110 CFU) of the same strain of Str uberis caused equally severe mastitis in 3 quarters colonized with C bovis and in 1 bacteriologically negative quarter in 2 cows. Streptococcus uberis was isolated from all inoculated quarters, and all quarters developed clinical mastitis by 2 days after inoculation. Two quarters colonized with C bovis and 2 bacteriologically negative quarters were inoculated once with 25 CFU and once with 240 CFU of a different strain of Str uberis (ATCC 27958). Streptococcus uberis was never isolated from inoculated quarters, and changes in milk yield or number of somatic cells were not observed.     

Susceptibility of Arcanobacterium pyogenes from different sources to tetracycline, macrolide and lincosamide antimicrobial agents

Chlortetracycline, oxytetracycline, and the macrolide, tylosin, are extensively used for growth promotion and disease prophylaxis in the cattle and swine industries in the US. Arcanobacterium pyogenes, a common inhabitant of the mucosal surfaces of cattle and swine, is also a pathogen associated with a variety of infections in these animals. A broth microdilution technique was used to determine the antimicrobial susceptibility of 48 A. pyogenes isolates to macrolides, lincosamides and tetracyclines. The MIC50 and MIC90 for chlortetracycline were 0.12 and 8 mg/l, respectively. Similarly, the MIC50 and MIC90 for oxytetracycline were 0.25 and 8 mg/l, while the MIC50 and MIC90 for tetracycline were 0.25 and 16 mg/l, respectively. The MIC50 and the MIC90 were < or = 0.06 and >64 mg/l, respectively, for erythromycin, tylosin and clindamycin. This resistance pattern indicated that some of these A. pyogenes isolates may carry an MLS(B) resistance determinant. A. pyogenes isolates (12.5%) were resistant to erythromycin, and this percentage doubled when MICs were performed following induction with erythromycin. Of the 48 A. pyogenes isolates, 25 and 41.7% were resistant to MLS(B) antimicrobial agents and the tetracycline derivatives, respectively. MLS(B) resistance was present in 22.2 and 35.3% of A. pyogenes isolates of bovine (n=27) or porcine (n=17) origin. In contrast, 70.6% of porcine isolates were resistant to the tetracyclines, compared with 25.9% of bovine isolates. These data suggest that a large proportion of A. pyogenes field isolates may be resistant to these commonly used antimicrobial agents.     

IgG antibody response to Corynebacterium pyogenes,Peptococcus indolicus and microaerophilic cocci in natural and experimental mastitis

A mixed inoculum of C. pyogenes, P. indolicus and a microaerophilic coccus was used to induce experimental mastitis in two heifers. During the course of the infection, the IgG titres against these bacteria were monitored by the indirect immunofluorescent antibody technique. At the same time, the course of the infection was studied bacteriologically and by determination of clinical-chemical parameters. In both heifers, high titres of IgG against C. pyogenes, P. indolicus and the microaerophilic coccus were detected. The reactivity of the IgG towards surface-associated antigens of these bacteria seemed to reside mainly in the IgG2 subclass. Sera obtained from Denmark from heifers with experimental mastitis, which had been induced similarly, were also found to have significantly raised IgG titres against C. pyogenes and P. indolicus. Sera from naturally infected heifers and dry cows, from whose udder secretions C. pyogenes and P. indolicus had been isolated, were examined comparatively with sera from healthy heifers and dry cows, respectively, for the presence of IgG against these two bacteria. Statistically significant differences in IgG titres against both bacteria were demonstrated between pregnant heifers with mastitis and healthy pregnant heifers and between dry cows with mastitis and clinically healthy controls.